Cell Counting:Article Title: Synergistic 3D-bioprinted scaffold with multi-level adaptability for vascularized bone regeneration via osteogenesis-angiogenesis coupling
Article Snippet: DMOG was purchased from APExBIO Technology LLC (Houston, USA). .. Calcein-AM/PI Cell Viability/Cytotoxicity Assay Kit, Cell-Counting Kit-8 (CCK-8) Kit, Alkaline Phosphatase Assay Kit, BCA Protein Assay Kit, Alizarin Red staining solution, Crystalline Violet staining solution, Penicillin/Streptomycin solution, and trypsin were obtained from Beyotime Biotechnology (Shanghai, China). .. DMEM/F12, DMEM and fetal bovine serum (FBS) was received from Gibco (Grand Island, USA).
ALP Assay:Article Title: Synergistic 3D-bioprinted scaffold with multi-level adaptability for vascularized bone regeneration via osteogenesis-angiogenesis coupling
Article Snippet: DMOG was purchased from APExBIO Technology LLC (Houston, USA). .. Calcein-AM/PI Cell Viability/Cytotoxicity Assay Kit, Cell-Counting Kit-8 (CCK-8) Kit, Alkaline Phosphatase Assay Kit, BCA Protein Assay Kit, Alizarin Red staining solution, Crystalline Violet staining solution, Penicillin/Streptomycin solution, and trypsin were obtained from Beyotime Biotechnology (Shanghai, China). .. DMEM/F12, DMEM and fetal bovine serum (FBS) was received from Gibco (Grand Island, USA).
Bicinchoninic Acid Protein Assay:Article Title: Synergistic 3D-bioprinted scaffold with multi-level adaptability for vascularized bone regeneration via osteogenesis-angiogenesis coupling
Article Snippet: DMOG was purchased from APExBIO Technology LLC (Houston, USA). .. Calcein-AM/PI Cell Viability/Cytotoxicity Assay Kit, Cell-Counting Kit-8 (CCK-8) Kit, Alkaline Phosphatase Assay Kit, BCA Protein Assay Kit, Alizarin Red staining solution, Crystalline Violet staining solution, Penicillin/Streptomycin solution, and trypsin were obtained from Beyotime Biotechnology (Shanghai, China). .. DMEM/F12, DMEM and fetal bovine serum (FBS) was received from Gibco (Grand Island, USA).
Staining:Article Title: Synergistic 3D-bioprinted scaffold with multi-level adaptability for vascularized bone regeneration via osteogenesis-angiogenesis coupling
Article Snippet: DMOG was purchased from APExBIO Technology LLC (Houston, USA). .. Calcein-AM/PI Cell Viability/Cytotoxicity Assay Kit, Cell-Counting Kit-8 (CCK-8) Kit, Alkaline Phosphatase Assay Kit, BCA Protein Assay Kit, Alizarin Red staining solution, Crystalline Violet staining solution, Penicillin/Streptomycin solution, and trypsin were obtained from Beyotime Biotechnology (Shanghai, China). .. DMEM/F12, DMEM and fetal bovine serum (FBS) was received from Gibco (Grand Island, USA).
Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy
Article Snippet: All DNA sequences used in this study were obtained from Sangon Biotech Co. Ltd. (Shanghai, China) and purified using HPLC ( ). .. The reagents and materials used in this chapter include: 3-Azidopropyl triphenylphosphonium bromide (Shanghai Aladdin Biotechnology Co., Ltd.); oligonucleotides (Shanghai Sangon Biotechnology Co., Ltd.); 0.25% trypsin cell digestion solution (C0201, Shanghai Beyotime Biotechnology Co., Ltd.); DMEM/F12 medium, DAPI staining solution (EE0011-A), enhanced CCK-8 assay kit (CT0001-B), Western primary antibody dilution (ED0013), Western secondary antibody dilution (ED0014), BCA protein quantification kit (EC0001-A), ultra-sensitive chemiluminescent kit (ED0015-B) (Shandong Sparkjade Biotechnology Co., Ltd.); 5 × TBE buffer (pH8.0), 20 × PBS buffer (pH7.4) (Shanghai Sangon Biotechnology Co., Ltd.); MgCl 2 (Macklin Biochemical); Tris-HCl, SYBR Green, Mito-Tracker Green Mitochondrial Probe, JC-1 Mitochondrial Membrane Potential Assay Kit (C2006), Reactive Oxygen Species Assay Kit (S0033S), PMSF (100 mM) phenylmethanesulfonyl fluoride, RIPA lysis buffer (P0013B), enhanced mitochondrial ATP assay kit (S0027) (Shanghai Beyotime Biotechnology Co., Ltd.); Malondialdehyde (MDA) content assay kit (AK288), Superoxide Dismutase (SOD) activity assay kit (AK060), Reduced Glutathione (GSH) content assay kit (AK102) (Beijing Solarbio Science & Technology Co., Ltd.); HO-1 specific antibody (Huabio, HA721428, 1:1000), NRF-2 specific antibody (Huabio, ET1705-511, 1:1000), cleaved-caspase 3 specific antibody (Huabio, ET1705-511, 1:1000), LC3-II specific antibody (Proteintech, 29932-1-AP, 1:1000), P62 specific antibody (Proteintech, 66,854-1-lg, 1:1000), COX4 specific antibody (Proteintech, YN1882, 1:1000), β-actin specific antibody, and the dual-marker autophagy adenovirus (HBAD-mRFP-GFP-LC3) (Beijing Solarbio Science & Technology Co., Ltd.). .. The instruments used in this chapter's experiments include the following: a centrifuge (Eppendorf, Germany); pipettes (Thermo, USA); a vortex mixer (Scilogex, USA); a TU-100 dry bath incubator (Yiheng Instruments, China); a magnetic stirrer (Shanghai Shangyi, China); a Nano-S90 particle size analyzer (Malvern, UK); an atomic force microscope (Bruker, Germany); a gel imaging and analysis system (Bio-Rad, USA); a small animal in vivo imaging system (Spectral Instruments, USA); a laser scanning confocal microscope (Zeiss, Germany); a Talos L120C G2 transmission electron microscope (Thermo Fisher Scientific, USA); a microplate reader (Bio-Rad, USA); an inverted microscope; a HE-120 multifunctional horizontal electrophoresis tank; a Bio-Rad electrophoresis system; a chemiluminescence imaging system; and a CO 2 incubator.
Single Cell:
Purification:
Isolation:Article Title: hUCMSC mitochondrial EVs confer neuroprotection after ischemia by Tom1l2-mediated mitochondrial fusion and Crls1–cardiolipin axis reprogramming
Article Snippet: .. Mitochondria were isolated from treated PC12 cells and fresh rat brain tissue using the Cell Mitochondrial Isolation Kit (Beyotime, Cat. No. C3601) and Tissue Mitochondrial Isolation Kit (Beyotime, Cat. No. C3606), respectively, according to the manufacturers’ instructions: Adherent PC12 cells were washed with pre-cooled PBS, digested with a trypsin–EDTA solution (Beyotime, Cat. No. C0201), and centrifuged (200× g , RT, 10 min) to collect pellets, which were subsequently washed by centrifugation (600× g , 4 °C, 5 min). ..
Centrifugation:Article Title: hUCMSC mitochondrial EVs confer neuroprotection after ischemia by Tom1l2-mediated mitochondrial fusion and Crls1–cardiolipin axis reprogramming
Article Snippet: .. Mitochondria were isolated from treated PC12 cells and fresh rat brain tissue using the Cell Mitochondrial Isolation Kit (Beyotime, Cat. No. C3601) and Tissue Mitochondrial Isolation Kit (Beyotime, Cat. No. C3606), respectively, according to the manufacturers’ instructions: Adherent PC12 cells were washed with pre-cooled PBS, digested with a trypsin–EDTA solution (Beyotime, Cat. No. C0201), and centrifuged (200× g , RT, 10 min) to collect pellets, which were subsequently washed by centrifugation (600× g , 4 °C, 5 min). ..
CCK-8 Assay:Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy
Article Snippet: All DNA sequences used in this study were obtained from Sangon Biotech Co. Ltd. (Shanghai, China) and purified using HPLC ( ). .. The reagents and materials used in this chapter include: 3-Azidopropyl triphenylphosphonium bromide (Shanghai Aladdin Biotechnology Co., Ltd.); oligonucleotides (Shanghai Sangon Biotechnology Co., Ltd.); 0.25% trypsin cell digestion solution (C0201, Shanghai Beyotime Biotechnology Co., Ltd.); DMEM/F12 medium, DAPI staining solution (EE0011-A), enhanced CCK-8 assay kit (CT0001-B), Western primary antibody dilution (ED0013), Western secondary antibody dilution (ED0014), BCA protein quantification kit (EC0001-A), ultra-sensitive chemiluminescent kit (ED0015-B) (Shandong Sparkjade Biotechnology Co., Ltd.); 5 × TBE buffer (pH8.0), 20 × PBS buffer (pH7.4) (Shanghai Sangon Biotechnology Co., Ltd.); MgCl 2 (Macklin Biochemical); Tris-HCl, SYBR Green, Mito-Tracker Green Mitochondrial Probe, JC-1 Mitochondrial Membrane Potential Assay Kit (C2006), Reactive Oxygen Species Assay Kit (S0033S), PMSF (100 mM) phenylmethanesulfonyl fluoride, RIPA lysis buffer (P0013B), enhanced mitochondrial ATP assay kit (S0027) (Shanghai Beyotime Biotechnology Co., Ltd.); Malondialdehyde (MDA) content assay kit (AK288), Superoxide Dismutase (SOD) activity assay kit (AK060), Reduced Glutathione (GSH) content assay kit (AK102) (Beijing Solarbio Science & Technology Co., Ltd.); HO-1 specific antibody (Huabio, HA721428, 1:1000), NRF-2 specific antibody (Huabio, ET1705-511, 1:1000), cleaved-caspase 3 specific antibody (Huabio, ET1705-511, 1:1000), LC3-II specific antibody (Proteintech, 29932-1-AP, 1:1000), P62 specific antibody (Proteintech, 66,854-1-lg, 1:1000), COX4 specific antibody (Proteintech, YN1882, 1:1000), β-actin specific antibody, and the dual-marker autophagy adenovirus (HBAD-mRFP-GFP-LC3) (Beijing Solarbio Science & Technology Co., Ltd.). .. The instruments used in this chapter's experiments include the following: a centrifuge (Eppendorf, Germany); pipettes (Thermo, USA); a vortex mixer (Scilogex, USA); a TU-100 dry bath incubator (Yiheng Instruments, China); a magnetic stirrer (Shanghai Shangyi, China); a Nano-S90 particle size analyzer (Malvern, UK); an atomic force microscope (Bruker, Germany); a gel imaging and analysis system (Bio-Rad, USA); a small animal in vivo imaging system (Spectral Instruments, USA); a laser scanning confocal microscope (Zeiss, Germany); a Talos L120C G2 transmission electron microscope (Thermo Fisher Scientific, USA); a microplate reader (Bio-Rad, USA); an inverted microscope; a HE-120 multifunctional horizontal electrophoresis tank; a Bio-Rad electrophoresis system; a chemiluminescence imaging system; and a CO 2 incubator.
Western Blot:Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy
Article Snippet: All DNA sequences used in this study were obtained from Sangon Biotech Co. Ltd. (Shanghai, China) and purified using HPLC ( ). .. The reagents and materials used in this chapter include: 3-Azidopropyl triphenylphosphonium bromide (Shanghai Aladdin Biotechnology Co., Ltd.); oligonucleotides (Shanghai Sangon Biotechnology Co., Ltd.); 0.25% trypsin cell digestion solution (C0201, Shanghai Beyotime Biotechnology Co., Ltd.); DMEM/F12 medium, DAPI staining solution (EE0011-A), enhanced CCK-8 assay kit (CT0001-B), Western primary antibody dilution (ED0013), Western secondary antibody dilution (ED0014), BCA protein quantification kit (EC0001-A), ultra-sensitive chemiluminescent kit (ED0015-B) (Shandong Sparkjade Biotechnology Co., Ltd.); 5 × TBE buffer (pH8.0), 20 × PBS buffer (pH7.4) (Shanghai Sangon Biotechnology Co., Ltd.); MgCl 2 (Macklin Biochemical); Tris-HCl, SYBR Green, Mito-Tracker Green Mitochondrial Probe, JC-1 Mitochondrial Membrane Potential Assay Kit (C2006), Reactive Oxygen Species Assay Kit (S0033S), PMSF (100 mM) phenylmethanesulfonyl fluoride, RIPA lysis buffer (P0013B), enhanced mitochondrial ATP assay kit (S0027) (Shanghai Beyotime Biotechnology Co., Ltd.); Malondialdehyde (MDA) content assay kit (AK288), Superoxide Dismutase (SOD) activity assay kit (AK060), Reduced Glutathione (GSH) content assay kit (AK102) (Beijing Solarbio Science & Technology Co., Ltd.); HO-1 specific antibody (Huabio, HA721428, 1:1000), NRF-2 specific antibody (Huabio, ET1705-511, 1:1000), cleaved-caspase 3 specific antibody (Huabio, ET1705-511, 1:1000), LC3-II specific antibody (Proteintech, 29932-1-AP, 1:1000), P62 specific antibody (Proteintech, 66,854-1-lg, 1:1000), COX4 specific antibody (Proteintech, YN1882, 1:1000), β-actin specific antibody, and the dual-marker autophagy adenovirus (HBAD-mRFP-GFP-LC3) (Beijing Solarbio Science & Technology Co., Ltd.). .. The instruments used in this chapter's experiments include the following: a centrifuge (Eppendorf, Germany); pipettes (Thermo, USA); a vortex mixer (Scilogex, USA); a TU-100 dry bath incubator (Yiheng Instruments, China); a magnetic stirrer (Shanghai Shangyi, China); a Nano-S90 particle size analyzer (Malvern, UK); an atomic force microscope (Bruker, Germany); a gel imaging and analysis system (Bio-Rad, USA); a small animal in vivo imaging system (Spectral Instruments, USA); a laser scanning confocal microscope (Zeiss, Germany); a Talos L120C G2 transmission electron microscope (Thermo Fisher Scientific, USA); a microplate reader (Bio-Rad, USA); an inverted microscope; a HE-120 multifunctional horizontal electrophoresis tank; a Bio-Rad electrophoresis system; a chemiluminescence imaging system; and a CO 2 incubator.
BIA-KA:Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy
Article Snippet: All DNA sequences used in this study were obtained from Sangon Biotech Co. Ltd. (Shanghai, China) and purified using HPLC ( ). .. The reagents and materials used in this chapter include: 3-Azidopropyl triphenylphosphonium bromide (Shanghai Aladdin Biotechnology Co., Ltd.); oligonucleotides (Shanghai Sangon Biotechnology Co., Ltd.); 0.25% trypsin cell digestion solution (C0201, Shanghai Beyotime Biotechnology Co., Ltd.); DMEM/F12 medium, DAPI staining solution (EE0011-A), enhanced CCK-8 assay kit (CT0001-B), Western primary antibody dilution (ED0013), Western secondary antibody dilution (ED0014), BCA protein quantification kit (EC0001-A), ultra-sensitive chemiluminescent kit (ED0015-B) (Shandong Sparkjade Biotechnology Co., Ltd.); 5 × TBE buffer (pH8.0), 20 × PBS buffer (pH7.4) (Shanghai Sangon Biotechnology Co., Ltd.); MgCl 2 (Macklin Biochemical); Tris-HCl, SYBR Green, Mito-Tracker Green Mitochondrial Probe, JC-1 Mitochondrial Membrane Potential Assay Kit (C2006), Reactive Oxygen Species Assay Kit (S0033S), PMSF (100 mM) phenylmethanesulfonyl fluoride, RIPA lysis buffer (P0013B), enhanced mitochondrial ATP assay kit (S0027) (Shanghai Beyotime Biotechnology Co., Ltd.); Malondialdehyde (MDA) content assay kit (AK288), Superoxide Dismutase (SOD) activity assay kit (AK060), Reduced Glutathione (GSH) content assay kit (AK102) (Beijing Solarbio Science & Technology Co., Ltd.); HO-1 specific antibody (Huabio, HA721428, 1:1000), NRF-2 specific antibody (Huabio, ET1705-511, 1:1000), cleaved-caspase 3 specific antibody (Huabio, ET1705-511, 1:1000), LC3-II specific antibody (Proteintech, 29932-1-AP, 1:1000), P62 specific antibody (Proteintech, 66,854-1-lg, 1:1000), COX4 specific antibody (Proteintech, YN1882, 1:1000), β-actin specific antibody, and the dual-marker autophagy adenovirus (HBAD-mRFP-GFP-LC3) (Beijing Solarbio Science & Technology Co., Ltd.). .. The instruments used in this chapter's experiments include the following: a centrifuge (Eppendorf, Germany); pipettes (Thermo, USA); a vortex mixer (Scilogex, USA); a TU-100 dry bath incubator (Yiheng Instruments, China); a magnetic stirrer (Shanghai Shangyi, China); a Nano-S90 particle size analyzer (Malvern, UK); an atomic force microscope (Bruker, Germany); a gel imaging and analysis system (Bio-Rad, USA); a small animal in vivo imaging system (Spectral Instruments, USA); a laser scanning confocal microscope (Zeiss, Germany); a Talos L120C G2 transmission electron microscope (Thermo Fisher Scientific, USA); a microplate reader (Bio-Rad, USA); an inverted microscope; a HE-120 multifunctional horizontal electrophoresis tank; a Bio-Rad electrophoresis system; a chemiluminescence imaging system; and a CO 2 incubator.
SYBR Green Assay:Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy
Article Snippet: All DNA sequences used in this study were obtained from Sangon Biotech Co. Ltd. (Shanghai, China) and purified using HPLC ( ). .. The reagents and materials used in this chapter include: 3-Azidopropyl triphenylphosphonium bromide (Shanghai Aladdin Biotechnology Co., Ltd.); oligonucleotides (Shanghai Sangon Biotechnology Co., Ltd.); 0.25% trypsin cell digestion solution (C0201, Shanghai Beyotime Biotechnology Co., Ltd.); DMEM/F12 medium, DAPI staining solution (EE0011-A), enhanced CCK-8 assay kit (CT0001-B), Western primary antibody dilution (ED0013), Western secondary antibody dilution (ED0014), BCA protein quantification kit (EC0001-A), ultra-sensitive chemiluminescent kit (ED0015-B) (Shandong Sparkjade Biotechnology Co., Ltd.); 5 × TBE buffer (pH8.0), 20 × PBS buffer (pH7.4) (Shanghai Sangon Biotechnology Co., Ltd.); MgCl 2 (Macklin Biochemical); Tris-HCl, SYBR Green, Mito-Tracker Green Mitochondrial Probe, JC-1 Mitochondrial Membrane Potential Assay Kit (C2006), Reactive Oxygen Species Assay Kit (S0033S), PMSF (100 mM) phenylmethanesulfonyl fluoride, RIPA lysis buffer (P0013B), enhanced mitochondrial ATP assay kit (S0027) (Shanghai Beyotime Biotechnology Co., Ltd.); Malondialdehyde (MDA) content assay kit (AK288), Superoxide Dismutase (SOD) activity assay kit (AK060), Reduced Glutathione (GSH) content assay kit (AK102) (Beijing Solarbio Science & Technology Co., Ltd.); HO-1 specific antibody (Huabio, HA721428, 1:1000), NRF-2 specific antibody (Huabio, ET1705-511, 1:1000), cleaved-caspase 3 specific antibody (Huabio, ET1705-511, 1:1000), LC3-II specific antibody (Proteintech, 29932-1-AP, 1:1000), P62 specific antibody (Proteintech, 66,854-1-lg, 1:1000), COX4 specific antibody (Proteintech, YN1882, 1:1000), β-actin specific antibody, and the dual-marker autophagy adenovirus (HBAD-mRFP-GFP-LC3) (Beijing Solarbio Science & Technology Co., Ltd.). .. The instruments used in this chapter's experiments include the following: a centrifuge (Eppendorf, Germany); pipettes (Thermo, USA); a vortex mixer (Scilogex, USA); a TU-100 dry bath incubator (Yiheng Instruments, China); a magnetic stirrer (Shanghai Shangyi, China); a Nano-S90 particle size analyzer (Malvern, UK); an atomic force microscope (Bruker, Germany); a gel imaging and analysis system (Bio-Rad, USA); a small animal in vivo imaging system (Spectral Instruments, USA); a laser scanning confocal microscope (Zeiss, Germany); a Talos L120C G2 transmission electron microscope (Thermo Fisher Scientific, USA); a microplate reader (Bio-Rad, USA); an inverted microscope; a HE-120 multifunctional horizontal electrophoresis tank; a Bio-Rad electrophoresis system; a chemiluminescence imaging system; and a CO 2 incubator.
Membrane:Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy
Article Snippet: All DNA sequences used in this study were obtained from Sangon Biotech Co. Ltd. (Shanghai, China) and purified using HPLC ( ). .. The reagents and materials used in this chapter include: 3-Azidopropyl triphenylphosphonium bromide (Shanghai Aladdin Biotechnology Co., Ltd.); oligonucleotides (Shanghai Sangon Biotechnology Co., Ltd.); 0.25% trypsin cell digestion solution (C0201, Shanghai Beyotime Biotechnology Co., Ltd.); DMEM/F12 medium, DAPI staining solution (EE0011-A), enhanced CCK-8 assay kit (CT0001-B), Western primary antibody dilution (ED0013), Western secondary antibody dilution (ED0014), BCA protein quantification kit (EC0001-A), ultra-sensitive chemiluminescent kit (ED0015-B) (Shandong Sparkjade Biotechnology Co., Ltd.); 5 × TBE buffer (pH8.0), 20 × PBS buffer (pH7.4) (Shanghai Sangon Biotechnology Co., Ltd.); MgCl 2 (Macklin Biochemical); Tris-HCl, SYBR Green, Mito-Tracker Green Mitochondrial Probe, JC-1 Mitochondrial Membrane Potential Assay Kit (C2006), Reactive Oxygen Species Assay Kit (S0033S), PMSF (100 mM) phenylmethanesulfonyl fluoride, RIPA lysis buffer (P0013B), enhanced mitochondrial ATP assay kit (S0027) (Shanghai Beyotime Biotechnology Co., Ltd.); Malondialdehyde (MDA) content assay kit (AK288), Superoxide Dismutase (SOD) activity assay kit (AK060), Reduced Glutathione (GSH) content assay kit (AK102) (Beijing Solarbio Science & Technology Co., Ltd.); HO-1 specific antibody (Huabio, HA721428, 1:1000), NRF-2 specific antibody (Huabio, ET1705-511, 1:1000), cleaved-caspase 3 specific antibody (Huabio, ET1705-511, 1:1000), LC3-II specific antibody (Proteintech, 29932-1-AP, 1:1000), P62 specific antibody (Proteintech, 66,854-1-lg, 1:1000), COX4 specific antibody (Proteintech, YN1882, 1:1000), β-actin specific antibody, and the dual-marker autophagy adenovirus (HBAD-mRFP-GFP-LC3) (Beijing Solarbio Science & Technology Co., Ltd.). .. The instruments used in this chapter's experiments include the following: a centrifuge (Eppendorf, Germany); pipettes (Thermo, USA); a vortex mixer (Scilogex, USA); a TU-100 dry bath incubator (Yiheng Instruments, China); a magnetic stirrer (Shanghai Shangyi, China); a Nano-S90 particle size analyzer (Malvern, UK); an atomic force microscope (Bruker, Germany); a gel imaging and analysis system (Bio-Rad, USA); a small animal in vivo imaging system (Spectral Instruments, USA); a laser scanning confocal microscope (Zeiss, Germany); a Talos L120C G2 transmission electron microscope (Thermo Fisher Scientific, USA); a microplate reader (Bio-Rad, USA); an inverted microscope; a HE-120 multifunctional horizontal electrophoresis tank; a Bio-Rad electrophoresis system; a chemiluminescence imaging system; and a CO 2 incubator.
Lysis:Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy
Article Snippet: All DNA sequences used in this study were obtained from Sangon Biotech Co. Ltd. (Shanghai, China) and purified using HPLC ( ). .. The reagents and materials used in this chapter include: 3-Azidopropyl triphenylphosphonium bromide (Shanghai Aladdin Biotechnology Co., Ltd.); oligonucleotides (Shanghai Sangon Biotechnology Co., Ltd.); 0.25% trypsin cell digestion solution (C0201, Shanghai Beyotime Biotechnology Co., Ltd.); DMEM/F12 medium, DAPI staining solution (EE0011-A), enhanced CCK-8 assay kit (CT0001-B), Western primary antibody dilution (ED0013), Western secondary antibody dilution (ED0014), BCA protein quantification kit (EC0001-A), ultra-sensitive chemiluminescent kit (ED0015-B) (Shandong Sparkjade Biotechnology Co., Ltd.); 5 × TBE buffer (pH8.0), 20 × PBS buffer (pH7.4) (Shanghai Sangon Biotechnology Co., Ltd.); MgCl 2 (Macklin Biochemical); Tris-HCl, SYBR Green, Mito-Tracker Green Mitochondrial Probe, JC-1 Mitochondrial Membrane Potential Assay Kit (C2006), Reactive Oxygen Species Assay Kit (S0033S), PMSF (100 mM) phenylmethanesulfonyl fluoride, RIPA lysis buffer (P0013B), enhanced mitochondrial ATP assay kit (S0027) (Shanghai Beyotime Biotechnology Co., Ltd.); Malondialdehyde (MDA) content assay kit (AK288), Superoxide Dismutase (SOD) activity assay kit (AK060), Reduced Glutathione (GSH) content assay kit (AK102) (Beijing Solarbio Science & Technology Co., Ltd.); HO-1 specific antibody (Huabio, HA721428, 1:1000), NRF-2 specific antibody (Huabio, ET1705-511, 1:1000), cleaved-caspase 3 specific antibody (Huabio, ET1705-511, 1:1000), LC3-II specific antibody (Proteintech, 29932-1-AP, 1:1000), P62 specific antibody (Proteintech, 66,854-1-lg, 1:1000), COX4 specific antibody (Proteintech, YN1882, 1:1000), β-actin specific antibody, and the dual-marker autophagy adenovirus (HBAD-mRFP-GFP-LC3) (Beijing Solarbio Science & Technology Co., Ltd.). .. The instruments used in this chapter's experiments include the following: a centrifuge (Eppendorf, Germany); pipettes (Thermo, USA); a vortex mixer (Scilogex, USA); a TU-100 dry bath incubator (Yiheng Instruments, China); a magnetic stirrer (Shanghai Shangyi, China); a Nano-S90 particle size analyzer (Malvern, UK); an atomic force microscope (Bruker, Germany); a gel imaging and analysis system (Bio-Rad, USA); a small animal in vivo imaging system (Spectral Instruments, USA); a laser scanning confocal microscope (Zeiss, Germany); a Talos L120C G2 transmission electron microscope (Thermo Fisher Scientific, USA); a microplate reader (Bio-Rad, USA); an inverted microscope; a HE-120 multifunctional horizontal electrophoresis tank; a Bio-Rad electrophoresis system; a chemiluminescence imaging system; and a CO 2 incubator.
ATP Assay:Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy
Article Snippet: All DNA sequences used in this study were obtained from Sangon Biotech Co. Ltd. (Shanghai, China) and purified using HPLC ( ). .. The reagents and materials used in this chapter include: 3-Azidopropyl triphenylphosphonium bromide (Shanghai Aladdin Biotechnology Co., Ltd.); oligonucleotides (Shanghai Sangon Biotechnology Co., Ltd.); 0.25% trypsin cell digestion solution (C0201, Shanghai Beyotime Biotechnology Co., Ltd.); DMEM/F12 medium, DAPI staining solution (EE0011-A), enhanced CCK-8 assay kit (CT0001-B), Western primary antibody dilution (ED0013), Western secondary antibody dilution (ED0014), BCA protein quantification kit (EC0001-A), ultra-sensitive chemiluminescent kit (ED0015-B) (Shandong Sparkjade Biotechnology Co., Ltd.); 5 × TBE buffer (pH8.0), 20 × PBS buffer (pH7.4) (Shanghai Sangon Biotechnology Co., Ltd.); MgCl 2 (Macklin Biochemical); Tris-HCl, SYBR Green, Mito-Tracker Green Mitochondrial Probe, JC-1 Mitochondrial Membrane Potential Assay Kit (C2006), Reactive Oxygen Species Assay Kit (S0033S), PMSF (100 mM) phenylmethanesulfonyl fluoride, RIPA lysis buffer (P0013B), enhanced mitochondrial ATP assay kit (S0027) (Shanghai Beyotime Biotechnology Co., Ltd.); Malondialdehyde (MDA) content assay kit (AK288), Superoxide Dismutase (SOD) activity assay kit (AK060), Reduced Glutathione (GSH) content assay kit (AK102) (Beijing Solarbio Science & Technology Co., Ltd.); HO-1 specific antibody (Huabio, HA721428, 1:1000), NRF-2 specific antibody (Huabio, ET1705-511, 1:1000), cleaved-caspase 3 specific antibody (Huabio, ET1705-511, 1:1000), LC3-II specific antibody (Proteintech, 29932-1-AP, 1:1000), P62 specific antibody (Proteintech, 66,854-1-lg, 1:1000), COX4 specific antibody (Proteintech, YN1882, 1:1000), β-actin specific antibody, and the dual-marker autophagy adenovirus (HBAD-mRFP-GFP-LC3) (Beijing Solarbio Science & Technology Co., Ltd.). .. The instruments used in this chapter's experiments include the following: a centrifuge (Eppendorf, Germany); pipettes (Thermo, USA); a vortex mixer (Scilogex, USA); a TU-100 dry bath incubator (Yiheng Instruments, China); a magnetic stirrer (Shanghai Shangyi, China); a Nano-S90 particle size analyzer (Malvern, UK); an atomic force microscope (Bruker, Germany); a gel imaging and analysis system (Bio-Rad, USA); a small animal in vivo imaging system (Spectral Instruments, USA); a laser scanning confocal microscope (Zeiss, Germany); a Talos L120C G2 transmission electron microscope (Thermo Fisher Scientific, USA); a microplate reader (Bio-Rad, USA); an inverted microscope; a HE-120 multifunctional horizontal electrophoresis tank; a Bio-Rad electrophoresis system; a chemiluminescence imaging system; and a CO 2 incubator.
Multiple Displacement Amplification:Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy
Article Snippet: All DNA sequences used in this study were obtained from Sangon Biotech Co. Ltd. (Shanghai, China) and purified using HPLC ( ). .. The reagents and materials used in this chapter include: 3-Azidopropyl triphenylphosphonium bromide (Shanghai Aladdin Biotechnology Co., Ltd.); oligonucleotides (Shanghai Sangon Biotechnology Co., Ltd.); 0.25% trypsin cell digestion solution (C0201, Shanghai Beyotime Biotechnology Co., Ltd.); DMEM/F12 medium, DAPI staining solution (EE0011-A), enhanced CCK-8 assay kit (CT0001-B), Western primary antibody dilution (ED0013), Western secondary antibody dilution (ED0014), BCA protein quantification kit (EC0001-A), ultra-sensitive chemiluminescent kit (ED0015-B) (Shandong Sparkjade Biotechnology Co., Ltd.); 5 × TBE buffer (pH8.0), 20 × PBS buffer (pH7.4) (Shanghai Sangon Biotechnology Co., Ltd.); MgCl 2 (Macklin Biochemical); Tris-HCl, SYBR Green, Mito-Tracker Green Mitochondrial Probe, JC-1 Mitochondrial Membrane Potential Assay Kit (C2006), Reactive Oxygen Species Assay Kit (S0033S), PMSF (100 mM) phenylmethanesulfonyl fluoride, RIPA lysis buffer (P0013B), enhanced mitochondrial ATP assay kit (S0027) (Shanghai Beyotime Biotechnology Co., Ltd.); Malondialdehyde (MDA) content assay kit (AK288), Superoxide Dismutase (SOD) activity assay kit (AK060), Reduced Glutathione (GSH) content assay kit (AK102) (Beijing Solarbio Science & Technology Co., Ltd.); HO-1 specific antibody (Huabio, HA721428, 1:1000), NRF-2 specific antibody (Huabio, ET1705-511, 1:1000), cleaved-caspase 3 specific antibody (Huabio, ET1705-511, 1:1000), LC3-II specific antibody (Proteintech, 29932-1-AP, 1:1000), P62 specific antibody (Proteintech, 66,854-1-lg, 1:1000), COX4 specific antibody (Proteintech, YN1882, 1:1000), β-actin specific antibody, and the dual-marker autophagy adenovirus (HBAD-mRFP-GFP-LC3) (Beijing Solarbio Science & Technology Co., Ltd.). .. The instruments used in this chapter's experiments include the following: a centrifuge (Eppendorf, Germany); pipettes (Thermo, USA); a vortex mixer (Scilogex, USA); a TU-100 dry bath incubator (Yiheng Instruments, China); a magnetic stirrer (Shanghai Shangyi, China); a Nano-S90 particle size analyzer (Malvern, UK); an atomic force microscope (Bruker, Germany); a gel imaging and analysis system (Bio-Rad, USA); a small animal in vivo imaging system (Spectral Instruments, USA); a laser scanning confocal microscope (Zeiss, Germany); a Talos L120C G2 transmission electron microscope (Thermo Fisher Scientific, USA); a microplate reader (Bio-Rad, USA); an inverted microscope; a HE-120 multifunctional horizontal electrophoresis tank; a Bio-Rad electrophoresis system; a chemiluminescence imaging system; and a CO 2 incubator.
Activity Assay:Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy
Article Snippet: All DNA sequences used in this study were obtained from Sangon Biotech Co. Ltd. (Shanghai, China) and purified using HPLC ( ). .. The reagents and materials used in this chapter include: 3-Azidopropyl triphenylphosphonium bromide (Shanghai Aladdin Biotechnology Co., Ltd.); oligonucleotides (Shanghai Sangon Biotechnology Co., Ltd.); 0.25% trypsin cell digestion solution (C0201, Shanghai Beyotime Biotechnology Co., Ltd.); DMEM/F12 medium, DAPI staining solution (EE0011-A), enhanced CCK-8 assay kit (CT0001-B), Western primary antibody dilution (ED0013), Western secondary antibody dilution (ED0014), BCA protein quantification kit (EC0001-A), ultra-sensitive chemiluminescent kit (ED0015-B) (Shandong Sparkjade Biotechnology Co., Ltd.); 5 × TBE buffer (pH8.0), 20 × PBS buffer (pH7.4) (Shanghai Sangon Biotechnology Co., Ltd.); MgCl 2 (Macklin Biochemical); Tris-HCl, SYBR Green, Mito-Tracker Green Mitochondrial Probe, JC-1 Mitochondrial Membrane Potential Assay Kit (C2006), Reactive Oxygen Species Assay Kit (S0033S), PMSF (100 mM) phenylmethanesulfonyl fluoride, RIPA lysis buffer (P0013B), enhanced mitochondrial ATP assay kit (S0027) (Shanghai Beyotime Biotechnology Co., Ltd.); Malondialdehyde (MDA) content assay kit (AK288), Superoxide Dismutase (SOD) activity assay kit (AK060), Reduced Glutathione (GSH) content assay kit (AK102) (Beijing Solarbio Science & Technology Co., Ltd.); HO-1 specific antibody (Huabio, HA721428, 1:1000), NRF-2 specific antibody (Huabio, ET1705-511, 1:1000), cleaved-caspase 3 specific antibody (Huabio, ET1705-511, 1:1000), LC3-II specific antibody (Proteintech, 29932-1-AP, 1:1000), P62 specific antibody (Proteintech, 66,854-1-lg, 1:1000), COX4 specific antibody (Proteintech, YN1882, 1:1000), β-actin specific antibody, and the dual-marker autophagy adenovirus (HBAD-mRFP-GFP-LC3) (Beijing Solarbio Science & Technology Co., Ltd.). .. The instruments used in this chapter's experiments include the following: a centrifuge (Eppendorf, Germany); pipettes (Thermo, USA); a vortex mixer (Scilogex, USA); a TU-100 dry bath incubator (Yiheng Instruments, China); a magnetic stirrer (Shanghai Shangyi, China); a Nano-S90 particle size analyzer (Malvern, UK); an atomic force microscope (Bruker, Germany); a gel imaging and analysis system (Bio-Rad, USA); a small animal in vivo imaging system (Spectral Instruments, USA); a laser scanning confocal microscope (Zeiss, Germany); a Talos L120C G2 transmission electron microscope (Thermo Fisher Scientific, USA); a microplate reader (Bio-Rad, USA); an inverted microscope; a HE-120 multifunctional horizontal electrophoresis tank; a Bio-Rad electrophoresis system; a chemiluminescence imaging system; and a CO 2 incubator.
ROS Assay:
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